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Epidemiological Survey On Gallibacterium Anatis Infection In Shanxi,shandong And Henan Province And Establishment And Application Of Random Amplified Polymorphic DNA For Gallibacterium Anatis Typing

Posted on:2011-11-03Degree:MasterType:Thesis
Country:ChinaCandidate:R Y FuFull Text:PDF
GTID:2143360308985340Subject:Prevention of Veterinary Medicine
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Layer oviduct cysts has been one of the diseases which troubles producers. The disease predominantly affects the respiratory tracts and reproductive tracts of layers, and the main clinic symptoms are drop in egg production, decline in quality, delay in production peak, deformation in eggshell, which lead to oviduct cysts forming penguin-like chicken as the main feature. It is reported that Gallibacterium anatis is associated with layer oviduct cysts. Bacteria belonging to genus Gallibacterium anatis seem to have a wide host spectrum, and cause a large economic losses in poultry industry.The serum epidemiology of the infection rate, species, age and other aspects of Gallibacterium anatis was investigated, from 1314 serum samples of 86 health chickens from different regions in Henan, Shandong and Shanxi province and 384 serum samples of diseased chickens from 12 farms in Henan province, which was detected by latex micro-agglutionation.The results showed that the infection rate of serum from 86 clinical healthy chickens reached 44.37% (583/1314), the infection rate of serotypeⅠof Gallibacterium anatis was1 1.80% (155/1314), serotypeⅡwas 22.98% (302/1314), serotypeⅣwas 9.60% (126/1314);The infection rate of serum from 12 farms in reached 48.4% (186/384);serotypeⅠof Gallibacterium anatis was 4.4% (17/384), serotypeⅡw as 21.4% (82/384), serotypeⅣwas 37.5% (144/384)。There were significant differences in infection rate of different breeds and ages of chickens, the infection rate of serum in Yukou was higher than Qingjiao chickens, serum infection rate of the puerperal was higher than the antepartum.Specific primers were designed and partial sequences of three housekeeping genes of rpoB, infB and atpD were amplified and sequenced from nine Gallibacteria anatis strains isolated in sequences of Gallibacteria anatis in the GenBank database. The seqences have been submitted to the GenBank database. Sequences of the three housekeeping genes of nine Gallibacteria strains were compared with that of reference strains of Avian Gallibacteria, Pasteurella and Actinobacillus. The results showed that the sequence homology was 98.1%~100% among Gallibacteria strains isolated in China, and 89.2%~97.1% Gallibacteria anatis strains in and out of China, and 82.1%~83.5% between Gallibacteria anatis strains and other genus strains of Pasteurella.The phylogenetic tree for three housekeeping genes was established,there were cluster of Gallibacteria anatis Chinese isolates,cluster of Gallibacteria anatis out of China and cluster of other genuses of pasteurella family. and the results indicated that the hereditary distance of Gallibacteria anatis was closed in China, the hereditary distance of Gallibacteria anatis in China was far from that out of China.In order to provide the reference for investigating the epidemic origin of Gallibacterium anatis, typing of Gallibacterium anatis and the relationship between Random Amplified Polymorphic DNA(RAPD) typing and serotyping of Gallibacterium anatis isolations. Nine strains of Gallibacterium anatis isolated from layer flocks in Henan were systematically classified by serotyping and by genotyping using randomly amplified polymorphic DNA (RAPD) with eight random primers, and three reference strains of Pasteurella multocida, Escherichia coli and Streptococcus suis. were used as control. The Gallibacterium anatis infection was mainly caused by serotypeⅠ(1/ 9), serotypeⅡ(3/ 9) and serotypeⅣ(5/9). In RAPD study, the results showed that three efficient primers were screened from eight random primers. The heredity distance of different strains were got by SPSS13.0 and the polygenetic tree among these strains was drawn. According to the heredity distance, eleven strains used in this study can be divided into 4 c1ustering groups, and nine strains of Gallibacterium anatis were divided into 4 c1ustering subgroups, three reference strains formed three c1ustering groups respectively. Gallibacterium anatis isolates of different serotype had similar finger print map, Gallibacterium anatis isolates of same serotype did not have the same finger print map. According to it, RAPD typing is one of better methods in mumerator epidemiological investigation,isolation and identification of Gallibacterium anatis,at present.
Keywords/Search Tags:Gallibacterium anatis, serotype, RAPD, Genotype, evolution analyse, epidemiologic survey
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